Probiotic Bacillus licheniformis MCC2514 and Bifidobacterium breve NCIM 5671 Regulates GATA3 and Foxp3 Expression in the Elevated Disease Condition.

TNBS-induced ulcerative colitis was evaluated using Bacillus licheniformis MCC 2514 (B. licheniformis) and Bifidobacterium breve NCIM 5671 (Bf. breve) as immune modulators. The study aims to analyze probiotic efficiency of ulcerative colitis induced by TNBS in Wistar rats. The tumor-like structure was found in the colon of TNBS inflammation-induced rats. Nitric oxide production was inhibited by about 65.2% fed with combination of bacteria and C-reactive protein, and decreased by 12% and 10.8% upon supplementing B. licheniformis and Bf. breve against the TNBS-treated rats, respectively. Liver damage was observed in the TNBS-treated rats; addition of probiotic bacteria reduced SGPT (75.4%) and SGOT (42.5%). On TNBS treatment, the transcriptional factor responsible for Th2 cell immune response (GATA3) was analyzed, and the elevation in gene expression (5.31-fold) was found. The FOXP-3 responsible for T-regulatory cells was expressed about 0.91-fold upon the treatment with a combination of bacteria. The expression of antioxidant genes such as iNOS (1.11-fold), GPx (1.29-fold), and PON1 (1.48-fold) has been increased when compared with that of the TNBS-treated group. The cytokines specific to Th2-driven immune response, such as IL-4, IL-5, and TNF-α, were reduced upon feeding the bacteria. It is observed that the B. licheniformis and Bf. breve used in the study have reduced Th2-driven immune response.

In vitro validation studies for adhesion factor and adhesion efficiency of probiotic Bacillus licheniformis MCC 2514 and Bifidobacterium breve NCIM 5671 on HT-29 cell lines.

Probiotic bacterial adhesion to the epithelial cell is a composite process and in vivo adhesion studies can be strengthened with the improved in vitro models for preliminary screening of potentially adherent strains. With this rationale, the study aimed is the first report to demonstrate the colonizing efficiency of probiotic Bacillus licheniformis MCC 2514 in comparison to Bifidobacterium breve NCIM 5671on HT-29 cell line. B. licheniformis (54.28 ± 0.99%) and Bif. breve (70.23 ± 0.85%) adhered in a higher percentage on fibronectin and mucin, respectively. However, the adhesion was higher for B. licheniformis when compared to Bif. breve. In adhesion score, B. licheniformis obtained about 138.85 ± 12.32, whereas Bif. breve got the score of 43.05 ± 9.12. The same trend continued in the adhesion percentage study, where B. licheniformis adhered 75.5 ± 5.2%, higher than Bif. breve which adhered 32.66 ± 3.2%. In invasion assay, both the bacteria significantly decreased the colonization of the pathogen Kocuria rhizophila ATCC 9341 about 97.32 ± 0.81% in the competitive assay, 97.87 ± 0.73% in exclusion assay and 82.19 ± 2.51% in displacement assay. The cytotoxicity effects of the test bacterial strains against HT-29 cell line through MTT assay determined no viability loss in the treated cells. Therefore, the data obtained from the in vitro studies showed that both B. licheniformis and Bif. breve had shown significantly good invasion on pathogen and adhesion capacity on HT-29 cell line.